@article{MoradianRochLendleinetal.2020,
  author    = {Moradian, Hanieh and Roch, Toralf and Lendlein, Andreas and Gossen, Manfred},
  title     = {mRNA transfection-induced activation of primary human monocytes and macrophages},
  series = {Scientific reports},
  volume    = {10},
  journal   = {Scientific reports},
  number    = {1},
  publisher = {Springer Nature},
  address   = {London},
  issn      = {2045-2322},
  doi       = {10.1038/s41598-020-60506-4},
  pages     = {1 -- 15},
  year      = {2020},
  abstract  = {Monocytes and macrophages are key players in maintaining immune homeostasis. Identifying strategies to manipulate their functions via gene delivery is thus of great interest for immunological research and biomedical applications. We set out to establish conditions for mRNA transfection in hard-to-transfect primary human monocytes and monocyte-derived macrophages due to the great potential of gene expression from in vitro transcribed mRNA for modulating cell phenotypes. mRNA doses, nucleotide modifications, and different carriers were systematically explored in order to optimize high mRNA transfer rates while minimizing cell stress and immune activation. We selected three commercially available mRNA transfection reagents including liposome and polymer-based formulations, covering different application spectra. Our results demonstrate that liposomal reagents can particularly combine high gene transfer rates with only moderate immune cell activation. For the latter, use of specific nucleotide modifications proved essential. In addition to improving efficacy of gene transfer, our findings address discrete aspects of innate immune activation using cytokine and surface marker expression, as well as cell viability as key readouts to judge overall transfection efficiency. The impact of this study goes beyond optimizing transfection conditions for immune cells, by providing a framework for assessing new gene carrier systems for monocyte and macrophage, tailored to specific applications.},
  language  = {en}
}
@article{MoradianGossenLendlein2022,
  author    = {Moradian, Hanieh and Gossen, Manfred and Lendlein, Andreas},
  title     = {Co-delivery of genes can be confounded by bicistronic vector design},
  series = {MRS Communications},
  volume    = {12},
  journal   = {MRS Communications},
  number    = {2},
  publisher = {Springer},
  address   = {Heidelberg},
  issn      = {2159-6859},
  doi       = {10.1557/s43579-021-00128-7},
  pages     = {145 -- 153},
  year      = {2022},
  abstract  = {Maximizing the efficiency of nanocarrier-mediated co-delivery of genes for co-expression in the same cell is critical for many applications. Strategies to maximize co-delivery of nucleic acids (NA) focused largely on carrier systems, with little attention towards payload composition itself. Here, we investigated the effects of different payload designs: co-delivery of two individual "monocistronic" NAs versus a single bicistronic NA comprising two genes separated by a 2A self-cleavage site. Unexpectedly, co-delivery via the monocistronic design resulted in a higher percentage of co-expressing cells, while predictive co-expression via the bicistronic design remained elusive. Our results will aid the application-dependent selection of the optimal methodology for co-delivery of genes.},
  language  = {en}
}
@article{MoradianRochAnthoferetal.2022,
  author    = {Moradian, Hanieh and Roch, Toralf and Anthofer, Larissa and Lendlein, Andreas and Gossen, Manfred},
  title     = {Chemical modification of uridine modulates mRNA-mediated proinflammatory and antiviral response in primary human macrophages},
  series = {Molecular therapy},
  volume    = {27},
  journal   = {Molecular therapy},
  publisher = {Cell Press},
  address   = {Cambridge},
  issn      = {2162-2531},
  doi       = {10.1016/j.omtn.2022.01.004},
  pages     = {854 -- 869},
  year      = {2022},
  abstract  = {In vitro transcribed (IVT)-mRNA has been accepted as a promising therapeutic modality. Advances in facile and rapid production technologies make IVT-mRNA an appealing alternative to protein- or virus-based medicines. Robust expression levels, lack of genotoxicity, and their manageable immunogenicity benefit its clinical applicability. We postulated that innate immune responses of therapeutically relevant human cells can be tailored or abrogated by combinations of 5'-end and internal IVT-mRNA modifications. Using primary human macrophages as targets, our data show the particular importance of uridine modifications for IVT-mRNA performance. Among five nucleotide modification schemes tested, 5-methoxy-uridine outperformed other modifications up to 4-fold increased transgene expression, triggering moderate proinflammatory and non-detectable antiviral responses. Macrophage responses against IVT-mRNAs exhibiting high immunogenicity (e.g., pseudouridine) could be minimized upon HPLC purification. Conversely, 5'-end modifications had only modest effects on mRNA expression and immune responses. Our results revealed how the uptake of chemically modified IVT-mRNA impacts human macrophages, responding with distinct patterns of innate immune responses concomitant with increased transient transgene expression. We anticipate our findings are instrumental to predictively address specific cell responses required for a wide range of therapeutic applications from eliciting controlled immunogenicity in mRNA vaccines to, e.g., completely abrogating cell activation in protein replacement therapies.},
  language  = {en}
}